CGRP receptor antagonists

ABSTRACT

The disclosures herein relate to novel compounds of formula 
                         
wherein Ar 1  and R 1  are as defined herein, and their use in treating, preventing, ameliorating, controlling or reducing cerebrovascular or vascular disorders associated with CGRP receptor function.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.15/336,893, filed on Oct. 28, 2016, which claims the benefit of GreatBritain Patent Application No. 1519194.3, filed Oct. 30, 2015. Both ofthe above-referenced patent applications are incorporated in theirentirety by reference herein.

TECHNICAL FIELD

This application relates to novel compounds and their use as CGRPreceptor antagonists. Compounds described herein may be useful in thetreatment or prevention of cerebrovascular or vascular disorders such asmigraine. The application is also directed to pharmaceuticalcompositions comprising these compounds and the manufacture and use ofthese compounds and compositions in the prevention or treatment of suchcerebrovascular or vascular disorders.

BACKGROUND OF THE INVENTION

Migraine is a highly disabling neurovascular disorder characterized byattacks of moderate to severe headache that are often associated withnausea, vomiting, photophobia, and phonophobia. The attacks can lastfrom 4 to 72 h, and the average attack frequency is 1 or 2 per month.About 20-30% of migraine patients experience transient focal neurologicsymptoms known as aura, which are usually visual and can precede oraccompany the headache. Migraine afflicts about 11% of adults worldwideand results in a significant socioeconomic burden, in terms of bothquality of life and lost productivity.

Whilst the pathomechanism of migraine is still unclear, one of theleading hypotheses is based on activation of the trigeminovascularsystem (TS). Several neuropeptides participate in this activation,calcitonin gene-related peptide (CGRP) playing a crucial role amongthem. CGRP exerts various biological effects through the peripheral andcentral nervous system (CNS). The functional CGRP-receptor (CGRP-R)complex has been well characterized, and novel therapeutic approachestarget CGRP itself and its receptors. This invention relates to thedevelopment of CGRP receptor antagonists (CGRP-RA).

CGRP, a 37-amino acid neuropeptide derived from the gene encodingcalcitonin, is formed from the alternative splicing of thecalcitonin/CGRP gene located on chromosome 11. In humans, CGRP has twoisoforms: α- and β-CGRP. The β-isoform differs from the α-isoform in theamino acids located at positions 3, 22 and 25. The chemical structure ofCGRP involves a disulphide bridge between residues 2 and 7 and anamidated C-terminus. The cyclic cysteine2-cysteine7 motif has a basicrole in receptor activation. In the human trigeminal ganglia (TRIG),CGRP-immunoreactive neurons account for up to 50% of all neurons. It hasbeen demonstrated through an in situ hybridization technique that 40% ofall nerve cell bodies contain CGRP mRNA and CGRP. Double immunostaininghas shown that in the human TRIG CGRP is co-localized with nitric oxidesynthase, substance P (SP), pituitary adenylate cyclase activatingpeptide (PACAP) and nociceptin, which may play a role in thepathomechanism of migraine.

The functional CGRP-R consists of three proteins: i) Calcitonin ReceptorLike Receptor (known as CRLR, CALCRL or CLR) is a seven-transmembranespanning protein, which forms the ligand binding site with; ii) RAMP1,determining the specificity of the receptor; and iii) the CGRP-Rcomponent protein (RCP) couples the receptor to intracellular signaltransduction pathways and to adenylyl cyclase.

It is thought that the C-terminal region of CGRP initially binds to thelarge N-terminal extracellular domain (ECD) of the receptor, likelymaking interactions with both CLR and RAMP1. This initial binding eventgreatly increases the local concentration of the N-terminal region ofCGRP in the vicinity of the juxtamembrane portion of CLR, allowing theirrelatively weak interaction to occur and resulting in receptoractivation. Since mutagenesis experiments indicated that most smallmolecule antagonists interacted with the ECD of CLR/RAMP1, it washypothesized that they bind to this region of the receptor and preventthe initial binding of CGRP to the receptor. A notable exception to thismodel of peptide binding and small molecule receptor antagonism is thehydroxypyridine class of antagonists, which apparently interact withtransmembrane domain 7 (TM7) in CLR and not with the extracellulardomain (Bell I M, J. Med. Chem., 2014, 57(19), 7838-58).

The first clinically tested CGRP-RA, olcegepant, was based on adipeptide backbone, had high molecular weight, and was not orallybioavailable. Nonetheless, when dosed intravenously, olcegepant provedto be an effective antimigraine agent, and this proof-of-concept studygreatly increased interest in the field. Following the success ofolcegepant, a number of orally acting CGRP-RAs were advanced to clinicaltrials. Telcagepant and compounds BI 44370, MK-3207, and BMS-927711 haveall been used for acute treatment of migraine as oral agents. Takentogether, the results from these clinical studies demonstrate thatCGRP-RAs can exhibit similar antimigraine efficacy to the gold standardtriptan drugs but with a significantly lower incidence of adverse eventsthan is typically observed with a triptan. It is worth noting that theavailable data indicate that these CGRP blockers do not causevasoconstriction and suggest that they may have a superiorcardiovascular safety profile to the triptans. One potential concernthat has been reported with some CGRP-RAs is the observation of elevatedlevels of liver transaminases in some patients, and this reportedly ledto the discontinuation of MK-3207. Although elevated liver enzymes werealso found in a small number of subjects after dosing with telcagepantfor an extended period, it is not clear if these findings are in someway mechanism-based or specific to these two compounds.

In clinical trials for acute migraine therapy, the CGRP-RAs displayedfavorable effects, but their frequent administration was associated withliver toxicity (the elevation of liver transaminases), which limitedtheir clinical use. Hence, there is a need to develop new CGRP-RAs whichdo not induce liver injury.

SUMMARY OF THE INVENTION

One possibility to address the risk of liver injury is to target anon-oral route of delivery for a small molecule which will place a lowerburden on the liver through first-pass exposure. The compounds of theinvention can be used for sub-cutaneous, intravenous and/or intranasalroutes of administration. The molecular profile for a CGRP-RA intendedfor such routes of administration differs from the profile required foran oral molecule: extremely high affinity and functional potency,coupled with extremely high solubility is required. Disclosed herein arenovel compounds, and the first medical use of said compounds as CGRPreceptor antagonists.

Compounds of the invention include compounds of formula (I)

or salts thereof, wherein R¹ is H or F and Ar¹ is an optionallysubstituted 5 membered heterocyclic ring containing at least twonitrogen atoms.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to novel compounds. The invention also relates tothe use of novel compounds as CGRP receptor antagonists. The inventionfurther relates to the use of compounds in the manufacture ofmedicaments for use as CGRP receptor antagonists. The invention furtherrelates to compounds, compositions and medicaments for the treatment ofcerebrovascular or vascular disorders such as migraine (includingsubtypes such as: migraine without aura, chronic migraine, puremenstrual migraine, menstrually-related migraine, migraine with aura,familial hemiplegic migraine, sporadic hemiplegic migraine, basilar-typemigraine, cyclical vomiting, abdominal migraine, benign paroxysmalvertigo of childhood, retinal migraine), status migrainosus, clusterheadache, dialysis headache, paroxysmal hemicrania, osteoarthritis, hotflashes associated with menopause or medically induced menopause due tosurgery or drug treatment, hemicrania continua, cyclic vomitingsyndrome, allergic rhinitis, or rosacea. The invention further relatesto compounds, compositions and medicaments for the treatment of broaderpain states and diseases involving neurogenic inflammation includingdental pain, earache, middle ear inflammation, sunburn, joint painassociated with osteoarthritis and rheumatoid arthritis, cancer pain,fibromyalgia, diabetic neuropathy, pain associated with inflammatorybowel disease—Crohn's disease, gout, complex regional pain syndrome,Behçet's disease, endometriosis pain, back pain or cough.

Compounds exemplified herein are based around the structure of formula(I):

or salts thereof, wherein R¹ is H or F and Ar¹ is an optionallysubstituted 5 membered heterocyclic ring containing at least twonitrogen atoms.In a particular embodiment, Ar¹ is an optionally substitutedfive-membered heterocyclic ring including at least two nitrogen atoms,wherein the optional substituents are selected from (C₁-C₆)alkyl, CO₂R²where R² is H or (C₁-C₃)alkyl.In a particular embodiment, R¹ is H.

In a more particular embodiment Ar¹ is a five-membered heterocyclic ringincluding two or three nitrogen atoms, optionally substituted with(C₁-C₆)alkyl.

In a particular embodiment, Ar¹ is selected from:

Further embodiments of the invention include methods of treatmentcomprising administering a compound of formulas (I) as a CGRP receptorantagonist. The treatment using a compound of formulas (I) may be in thetreatment of cerebrovascular disorders such as migraine (includingsubtypes such as: migraine without aura, chronic migraine, puremenstrual migraine, menstrually-related migraine, migraine with aura,familial hemiplegic migraine, sporadic hemiplegic migraine, basilar-typemigraine, cyclical vomiting, abdominal migraine, benign paroxysmalvertigo of childhood, retinal migraine), status migrainosus, clusterheadache, dialysis headache, paroxysmal hemicrania, osteoarthritis, hotflashes associated with menopause or medically induced menopause due tosurgery or drug treatment, hemicrania continua, cyclic vomitingsyndrome, allergic rhinitis, or rosacea. The invention further relatesto compounds, compositions and medicaments for the treatment of broaderpain states and diseases involving neurogenic inflammation includingdental pain, earache, middle ear inflammation, sunburn, joint painassociated with osteoarthritis and rheumatoid arthritis, cancer pain,fibromyalgia, diabetic neuropathy, pain associated with inflammatorybowel disease—Crohn's disease, gout, complex regional pain syndrome,Behçet's disease, endometriosis pain, back pain or cough.

Certain novel compounds of the invention show particularly highactivities as CGRP receptor antagonists. Exemplary compounds include:

The NMR and LCMS properties as well as the biological activities ofthese compounds are set out in Tables 2 and 3.

To the extent that any of the compounds described have chiral centres,the present invention extends to all optical isomers of such compounds,whether in the form of racemates or resolved enantiomers. The inventiondescribed herein relates to all crystal forms, solvates and hydrates ofany of the disclosed compounds however so prepared. To the extent thatany of the compounds and intermediates disclosed herein have acid orbasic centres such as carboxylates or amino groups, then all salt formsof said compounds are included herein. In the case of pharmaceuticaluses, the salt should be seen as being a pharmaceutically acceptablesalt.

Pharmaceutically acceptable salts that may be mentioned include acidaddition salts and base addition salts. Such salts may be formed byconventional means, for example by reaction of a free acid or a freebase form of a compound with one or more equivalents of an appropriateacid or base, optionally in a solvent, or in a medium in which the saltis insoluble, followed by removal of said solvent, or said medium, usingstandard techniques (e.g. in vacuo, by freeze-drying or by filtration).Salts may also be prepared by exchanging a counter-ion of a compound inthe form of a salt with another counter-ion, for example using asuitable ion exchange resin.

Examples of pharmaceutically acceptable salts include acid additionsalts derived from mineral acids and organic acids, and salts derivedfrom metals such as sodium, magnesium, or preferably, potassium andcalcium.

Examples of acid addition salts include acid addition salts formed withacetic, 2,2-dichloroacetic, adipic, alginic, aryl sulfonic acids (e.g.benzenesulfonic, naphthalene-2-sulfonic, naphthalene-1,5-disulfonic andp-toluenesulfonic), ascorbic (e.g. L-ascorbic), L-aspartic, benzoic,4-acetamidobenzoic, butanoic, (+)-camphoric, camphor-sulfonic,(+)-(1S)-camphor-10-sulfonic, capric, caproic, caprylic, cinnamic,citric, cyclamic, dodecyl sulfuric, ethane-1,2-disulfonic,ethanesulfonic, 2-hydroxyethanesulfonic, formic, fumaric, galactaric,gentisic, glucoheptonic, gluconic (e.g. D-gluconic), glucuronic (e.g.D-glucuronic), glutamic (e.g. L-glutamic), α-oxoglutaric, glycolic,hippuric, hydrobromic, hydrochloric, hydriodic, isethionic, lactic (e.g.(+)-L-lactic and (±)-DL-lactic), lactobionic, maleic, malic (e.g.(−)-L-malic), malonic, (±)-DL-mandelic, metaphosphoric, methanesulfonic,1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic,palmitic, pamoic, phosphoric, propionic, L-pyroglutamic, salicylic,4-amino-salicylic, sebacic, stearic, succinic, sulfuric, tannic,tartaric (e.g. (+)-L-tartaric), thiocyanic, undecylenic and valericacids.

Particular examples of salts are salts derived from mineral acids suchas hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric andsulfuric acids; from organic acids, such as tartaric, acetic, citric,malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic,arylsulfonic, pamoic acids; and from metals such as sodium, magnesium,or preferably, potassium and calcium.

Also encompassed are any solvates of the compounds and their salts.Preferred solvates are solvates formed by the incorporation into thesolid state structure (e.g. crystal structure) of the compounds of theinvention of molecules of a non-toxic pharmaceutically acceptablesolvent (referred to below as the solvating solvent). Examples of suchsolvents include water, alcohols (such as ethanol, isopropanol andbutanol) and dimethylsulfoxide. Solvates can be prepared byrecrystallising the compounds of the invention with a solvent or mixtureof solvents containing the solvating solvent. Whether or not a solvatehas been formed in any given instance can be determined by subjectingcrystals of the compound to analysis using well known and standardtechniques such as thermogravimetric analysis (TGE), differentialscanning calorimetry (DSC) and X-ray crystallography.

The solvates can be stoichiometric or non-stoichiometric solvates.Particular solvates may be hydrates, and examples of hydrates includehemihydrates, monohydrates and dihydrates.

For a more detailed discussion of solvates and the methods used to makeand characterise them, see Bryn et al., Solid-State Chemistry of Drugs,Second Edition, published by SSCI, Inc of West Lafayette, Ind., USA,1999, ISBN 0-967-06710-3.

“Pharmaceutically functional derivatives” of compounds as defined hereinincludes ester derivatives and/or derivatives that have, or provide for,the same biological function and/or activity as any relevant compound ofthe invention. Thus, for the purposes of this invention, the term alsoincludes prodrugs of compounds as defined herein.

The term “prodrug” of a relevant compound includes any compound that,following oral or parenteral administration, is metabolised in vivo toform that compound in an experimentally-detectable amount, and within apredetermined time (e.g. within a dosing interval of between 6 and 24hours (i.e. once to four times daily)).

Prodrugs of compounds may be prepared by modifying functional groupspresent on the compound in such a way that the modifications arecleaved, in vivo when such prodrug is administered to a mammaliansubject. The modifications typically are achieved by synthesizing theparent compound with a prodrug substituent. Prodrugs include compoundswherein a hydroxyl, amino, sulfhydryl, carboxyl or carbonyl group in acompound is bonded to any group that may be cleaved in vivo toregenerate the free hydroxyl, amino, sulfhydryl, carboxyl or carbonylgroup, respectively.

Examples of prodrugs include, but are not limited to, esters andcarbamates of hydroxyl functional groups, ester groups of carboxylfunctional groups, N-acyl derivatives and N-Mannich bases. Generalinformation on prodrugs may be found e.g. in Bundegaard, H. “Design ofProdrugs” p. 1-92, Elsevier, New York-Oxford (1985).

Definitions

C₁-C₆ Alkyl

Alkyl means an aliphatic hydrocarbon group. The alkyl group may bestraight or branched. “Branched” means that at least one carbon branchpoint is present in the group, for example isopropyl or tertiarybutyl.C₁-C₃ alkyl groups include methyl, ethyl, n-propyl, i-propyl. The alkylgroup may be optionally substituted.

Heterocyclic

Heterocyclic means a cyclic group which may be aromatic in which atleast one ring member is other than carbon. For example, at least onering member (for example one, two or three ring members) may be selectedfrom nitrogen, oxygen and sulphur. The point of attachment of heteroarylgroups may be via any atom of the ring system. Exemplary heteroarylgroups include indazolyl, imidazolyl, 1,2,4-triazolyl,quinolin-2(1H)-one, piperidinyl, and the like. Where the heteroarylgroup is 5-membered, the heteroaryl group includes imidazolyl,1,2,4-triazolyl, and the like.

Optionally Substituted

“Optionally substituted” as applied to any group means that the saidgroup may if desired be substituted with one or more substituents, whichmay be the same or different.

The term “pharmaceutical composition” in the context of this inventionmeans a composition comprising an active agent and comprisingadditionally one or more pharmaceutically acceptable carriers. Thecomposition may further contain ingredients selected from, for example,diluents, adjuvants, excipients, vehicles, preserving agents, fillers,disintegrating agents, wetting agents, emulsifying agents, suspendingagents, sweetening agents, flavouring agents, perfuming agents,antibacterial agents, antifungal agents, lubricating agents anddispersing agents, depending on the nature of the mode of administrationand dosage forms. The compositions may take the form, for example, oftablets, dragees, powders, elixirs, syrups, liquid preparationsincluding suspensions, sprays, inhalants, tablets, lozenges, emulsions,solutions, cachets, granules, capsules and suppositories, as well asliquid preparations for injections, including liposome preparations.

The dosages may be varied depending upon the requirements of thepatient, the severity of the condition being treated, and the compoundbeing employed. Determination of the proper dosage for a particularsituation is within the skill of the art. Generally, treatment isinitiated with the smaller dosages which are less than the optimum doseof the compound. Thereafter the dosage is increased by small incrementsuntil the optimum effect under the circumstances is reached. Forconvenience, the total daily dosage may be divided and administered inportions during the day if desired.

The magnitude of an effective dose of a compound will, of course, varywith the nature of the severity of the condition to be treated and withthe particular compound and its route of administration. The selectionof appropriate dosages is within the ability of one of ordinary skill inthis art, without undue burden. In general, the daily dose range may befrom about 10 μg to about 30 mg per kg body weight of a human andnon-human animal, preferably from about 50 μg to about 30 mg per kg ofbody weight of a human and non-human animal, for example from about 50μg to about 10 mg per kg of body weight of a human and non-human animal,for example from about 100 μg to about 30 mg per kg of body weight of ahuman and non-human animal, for example from about 100 μg to about 10 mgper kg of body weight of a human and non-human animal and mostpreferably from about 100 μg to about 1 mg per kg of body weight of ahuman and non-human animal.

Preparation of the Compounds of the Invention

Compounds of the invention may be prepared by routes including those inScheme 1. Details of many of the standard transformations such as thosein the routes below and others which could be used to perform the sametransformations can be found in standard reference textbooks such as“Organic Synthesis”, M. B. Smith, McGraw-Hill (1994) or “AdvancedOrganic Chemistry”, 4^(th) edition, J. March, John Wiley & Sons (1992).

Urea formations between amino acid intermediates, for example methylesters of amino acids, and amine intermediates can be formed underconditions using a coupling agent such as DSC or CDI in the presence ofa base such as triethylamine or DIPEA in solvents such as DMF and/orDCM. The methyl ester portion of the subsequently formed ureaderivatives can be saponified using aqueous bases such as lithiumhydroxide in a suitable solvent such as THF, MeOH, 1,4-dioxane, or amixture thereof. The acid intermediates thus formed can be convertedinto amide examples under standard conditions, for example using acoupling agent such as HATU, in the presence of a base such as DIPEA ortriethylamine in a suitable solvent such as DMF. The amine partners forsuch amide couplings can be prepared using an appropriate combination ofstandard transformations (for example reductive aminations using anamine, an aldehyde or ketone, and a reducing agent such as sodiumtriacetoxyborohydride or sodium cyanoborohydride, in a solvent such asMeOH or DCE, optionally in the presence of an additive such as aceticacid or zinc chloride; or alkylation using an alkyl halide and a strongbase such as sodium hydride in a suitable solvent such as DMF).Following standard transformations such as the above, or during such asequence of such transformations, removal of standard protecting groupsmay be necessary and can be undertaken using conditions which can befound in reference textbooks, for example “Protecting Groups”, 3^(rd)edition, P. J. Kocieński, Georg Thieme Verlag (2005). One suchtransformation is the removal of a tert-butoxycarbonyl group (commonlyknown as a Boc group) from an amine under acidic conditions such as HClin a solvent such as 1,4-dioxane, MeOH, EtOH, DCM or combinationsthereof. It can be appreciated that Boc deprotection of amineintermediates of the invention which possess additional basic centresmay result in hydrochloride salts of different stoichiometries. Forexample the Boc deprotection of an intermediate with one additionalbasic centre will result in the formation of a new amine intermediatewhich is for example the mono-hydrochloride or di-hydrochloride salt,which will often be used without neutralisation of the hydrochloridesalt to produce the free base of the intermediate, as it can beappreciated that in the subsequent amide formation an excess of a basesuch as DIPEA or triethylamine is typically used to neutralise thehydrochloride salt. Amine intermediates of the invention formed byBoc-deprotection which are used without neutralisation to the free baseare named herein as the hydrochloride (x HCl), and the present inventionextends to all salt forms of the said intermediates. Examples of theinvention may be transformed into further examples using standardtransformations such as those detailed above, for example saponificationof an ester using conditions such as those detailed above.

General Procedures

Where no preparative routes are included, the relevant intermediate iscommercially available. Commercial reagents were utilized withoutfurther purification. Room temperature (rt) refers to approximately20-27° C. ¹H NMR spectra were recorded at 400 MHz or 500 MHz on Bruker,Varian or JEOL instruments. Chemical shift values are expressed in partsper million (ppm), i.e. (δ)-values. The following abbreviations are usedfor the multiplicity of the NMR signals: s=singlet, br=broad, d=doublet,t=triplet, q=quartet, quin=quintet, h=heptet, dd=doublet of doublets,dt=double of triplets, m=multiplet. Coupling constants are listed as Jvalues, measured in Hz. NMR and mass spectroscopy results were correctedto account for background peaks. Where complex NMR spectra ofintermediates or examples exist due to the presence of tautomeric formsdata are provided for the major form observed. Chromatography refers tocolumn chromatography performed using silica and executed under positivepressure (flash chromatography) conditions. LCMS experiments werecarried out using electrospray conditions under the conditions below.LCMS data are given in the format: Mass ion, electrospray mode (positiveor negative), retention time (experimental text and Table 1); Mass ion,electrospray mode (positive or negative), retention time, approximatepurity (Table 2).

Method A.

Instruments: Hewlett Packard 1100 with G1315A DAD, Micromass ZQ; Column:Waters X-Bridge C-18, 2.5 micron, 2.1×20 mm or Phenomenex Gemini-NXC-18, 3 micron, 2.0×30 mm; Gradient [time (min)/solvent D in C (%)]:0.00/2, 0.10/2, 8.40/95, 10.00/95; Solvents: solvent C=2.5 L H₂O+2.5 mL28% ammonia in water solution; solvent D=2.5 L MeCN+135 mL H₂O+2.5 mL28% ammonia in water solution; Injection volume 1 μL; UV detection 230to 400 nM; column temperature 45° C.; Flow rate 1.5 mL/min.

Method B.

Instruments: Agilent Technologies 1260 Infinity LC with Chemstationsoftware, Diode Array Detector, Agilent 6120B Single Quadrupole MS withAPI-ES Source; Column: Phenomenex Gemini-NX C-18, 3 micron, 2.0×30 mm;Gradient [time (min)/solvent D in C (%)]: 0.00/5, 2.00/95, 2.50/95,2.60/5, 3.00/5; Solvents C and D are as described above in Method A;Injection volume 0.5 μL; UV detection 190 to 400 nM; column temperature40° C.; Flow rate 1.5 mL/min.

Method C.

Instruments: Waters Acquity H Class, Photo Diode Array, SQ Detector;Column: BEH C18, 1.7 micron, 2.1×50 mm; Gradient [time (min)/solvent Bin A (%)]: 0.00/5, 0.40/5, 0.8/35, 1.20/55, 2.50/100, 3.30/100 4.00/5;Solvents: solvent A=5 mM ammonium acetate and 0.1% formic acid in H₂O;solvent B=0.1% formic acid in MeCN; Injection volume 2 μL; UV detection200 to 400 nM; Mass detection 100 to 1200 AMU (+ve electrospray); columnat ambient temperature; Flow rate 0.5 mL/min.

Method D.

Instruments: Waters Acquity H Class, Photo Diode Array, SQ Detector;Column: X-Bridge C18, 5 micron, 150×4.6 mm; Gradient [time (min)/solventE in F (%)]: 0.01/10, 5.00/90, 7.00/100, 11.00/100, 11.01/10 12.00/10;Solvents: solvent E=0.1% ammonia in H₂O; solvent F=0.1% ammonia in MeCN;Injection volume 10 μL; UV detection 200 to 400 nM; Mass detection 60 to1000 AMU (+ve electrospray); column at ambient temperature; Flow rate1.0 mL/min.

Method E.

Instruments: Acquity UPLC coupled with SQD mass spectrometer; Column:Acquity UPLC BEH C18, 1.7 micron, 2.1×50 mm; Gradient [time(min)/solvent B in A (%)]: 0.00/5, 1.50/5, 8.75/80, 9.50/90, 9.80/90,12.00/5; Solvents: solvent A=10 mM aqueous solution of NH₄HCO₃ (adjustedto pH 10 with ammonia); solvent B=MeCN; Injection volume 2 μL; UVdetection 210 to 350 nM; column temperature 40° C.; Flow rate 0.9mL/min.

Abbreviations

-   CDI=1,1′-carbonyldiimidazole-   DCE=1,2-dichloroethane-   DCM=dichloromethane-   DIPEA=N,N-diisopropylethylamine-   DMAC=N,N-dimethylacetamide-   DMF=dimethylformamide-   DSC=N,N′-disuccinimidyl carbonate-   DMSO=dimethylsulfoxide-   ES=electrospray-   EtOAc=ethyl acetate-   h=hour(s)-   HATU=1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxid hexafluorophosphate-   L=liter-   LC=liquid chromatography-   LCMS=liquid chromatography mass spectrometry-   MeCN=acetonitrile-   min=minute(s)-   MS=mass spectrometry-   NMR=nuclear magnetic resonance-   rcf=relative centrifugal force-   rpm=revolutions per minute-   rt=room temperature-   s=second(s)-   THF=tetrahydrofuran-   TLC=thin-layer chromatography

Prefixes n-, s-, i-, t- and tert- have their usual meanings: normal,secondary, iso, and tertiary.

SYNTHESIS OF INTERMEDIATES Preparation of Carboxylic Acid IntermediatesTypical procedure for the preparation of carboxylic acid intermediatesvia urea formation and subsequent saponification, as exemplified by thepreparation of Intermediate 4,(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoicacid

Step 1) To a solution of (R)-methyl 2-amino-3-(7-methyl-1H-indazol-5-yl)propanoate (Intermediate 3, 6.05 g, 25.9 mmol) in DMF (60 mL) under N₂at approximately −20° C. was added CDI (8.40 g, 51.8 mmol) and themixture was stirred for 15 min while keeping the temperature below −10°C. A solution of H₂O (2.34 mL) in a few mL of DMF was added and stirringcontinued for 15 min while keeping the temperature below −10° C.3-(Piperidin-4-yl) quinolin-2(1H)-one (Intermediate 1, 6.99 g, 30.6mmol), DIPEA (4.93 mL, 28.2 mmol) and DCM (20 mL) were then added inthat order and the mixture was heated to 40° C. under N₂ for 12 h. Aftercooling to rt, 2M HCl (aq) (38.7 mL) was added and the mixture wasextracted twice with DCM. The combined organic extracts were washedthree times with H₂O, dried (Na₂SO₄) and concentrated in vacuo.Purification by flash chromatography, eluting with MeOH/DCM (5:95),yieldedmethyl(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoate(10.4 g, 21.3 mmol) as a light tan solid.

¹H NMR: (400 MHz, CDCl₃) δ: 1.40-1.60 (m, 2H), 1.95-1.97 (m, 2H), 2.46(s, 3H), 2.90-3.00 (m, 2H), 3.11-3.26 (m, 3H), 3.76 (s, 3H), 4.07-4.12(m, 2H), 4.86-4.91 (m, 1H), 5.18 (d, J=7.6, 1H), 6.93 (s, 1H), 7.17-7.21(m, 1H), 7.24 (s, 1H), 7.32 (s, 1H), 7.43-7.54 (m, 3H), 7.95 (s, 1H),10.70 (s, 2H).

Step 2) To a solution ofmethyl(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoate(9.79 g, 20.1 mmol) in 1,4-dioxane (150 mL) was added a solution ofLiOH.H₂O (1.26 g, 30.0 mmol) in H₂O (150 mL) and the mixture was stirredat rt for 2 h. The reaction mixture was concentrated in vacuo tonear-dryness and re-dissolved in H₂O before being acidified with aqueous2M HCl (approximately 15 mL) whilst being rapidly stirred. The resultingthick white precipitate was isolated by filtration and washed with H₂Ountil the washings were near neutral pH. Drying in vacuo yielded thetitle compound (8.11 g, 17.1 mmol) as an off-white solid.

Data in Table 1.

Intermediate 5,(2R)-2-({[4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)-3-(7-methyl-1H-indazol-5-yl)propanoicacid

Step 1) Et₃N (1.25 mL, 9.0 mmol) was added to a solution of (R)-methyl2-amino-3-(7-methyl-1H-indazol-5-yl)propanoate (Intermediate 3, 700 mg,3.0 mmol) and DSC (845 mg, 3.3 mmol) in DMF (20 mL) and the mixturestirred at rt for 30 min. 7-Fluoro-3-(piperidin-4-yl)quinolin-2(1H)-onehydrochloride (Intermediate 2, 933 mg, 3.3 mmol) was then addedportionwise and the reaction mixture stirred at rt overnight beforeconcentration in vacuo. The residue was partitioned between H₂O and DCM,with a small amount of MeOH added to aid dissolution, and the organicphase with washed with H₂O. After concentration in vacuo the residue waspurified by flash chromatography, eluting with EtOAc in MeOH (20:1), toyieldmethyl(2R)-2-({[4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)-3-(7-methyl-1H-indazol-5-yl)propanoate(462 mg, 0.91 mmol) as a yellow solid.

LCMS (Method A): m/z 506.3 (ES+), at 3.35 min.

¹H NMR: (400 MHz, DMSO-d₆) δ: ppm 1.22-1.37 (m, 2H), 1.73 (t, J=10.2,2H), 2.47 (s, 3H), 2.64-2.80 (m, 2H), 2.82-2.94 (m, 1H), 2.95-3.11 (m,2H), 3.59 (s, 3H), 4.08 (d, J=12.5, 2H), 4.21-4.33 (m, 1H), 6.85 (d,J=7.8, 1H), 6.97-7.10 (m, 3H), 7.41 (s, 1H), 7.59 (s, 1H), 7.70 (dd,J=8.2, 6.2, 1H), 7.87-8.10 (m, 1H), 11.85 (s, 1H), 13.04 (s, 1H)

Step 2)Methyl(2R)-2-({[4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)-3-(7-methyl-1H-indazol-5-yl)propanoate(462 mg, 0.91 mmol) was dissolved in THF (6 mL) and MeOH (1.2 mL) and anaqueous solution of LiOH (1M, 1.82 mL, 1.82 mmol) was added dropwise.After stirring at rt for 4 h the reaction mixture was concentrated undera stream of nitrogen, the residue dissolved in a minimum volume of H₂Oand acidified with 1M HCl. The resulting precipitate was isolated byfiltration, washed with cold H₂O and Et₂O to yield the title compound(406 mg, 0.83 mmol) as a pale yellow solid.

Data in Table 1.

Preparation of Amine Intermediates Intermediate 8,4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine hydrochloride

Step 1) tert-Butyl 4-oxopiperidine-1-carboxylate (Intermediate 7, 2.09g, 10.5 mmol) was added to a suspension of4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine (Intermediate 6, 1.66 g,10.0 mmol) in DCE (60 mL). The mixture was stirred at rt for 30 minbefore the addition of sodium triacetoxyborohydride (2.97 g, 14.0 mmol).After stirring at rt overnight tert-butyl 4-oxopiperidine-1-carboxylate(Intermediate 7, 210 mg, 1.05 mmol) was added and the mixture stirredfor 8 h at rt, followed by the addition of tert-butyl4-oxopiperidine-1-carboxylate (Intermediate 7, 600 mg, 3.01 mmol) andsodium triacetoxyborohydride (900 mg, 4.25 mmol) and stirring at rt for4 d. H₂O (100 mL) was added, the mixture was stirred for 2 min beforethe phases were separated and the aqueous layer (adjusted to pH 4.7) waswashed with DCM (2×30 mL). DCM (30 mL) was added to the aqueous phase,and the aqueous phase was adjusted to pH 7 by the addition of 1M NaOHbefore extraction with DCM (7×30 mL), adjusting the aqueous phase pH to7 before each extraction. The aqueous phase was further extracted withDCM (5×30 mL), adjusting the pH to 7.5 before each extraction. Thecombined organic phases were dried (Na₂SO₄), filtered and concentratedin vacuo to yield tert-butyl4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate(2.27 g, 6.50 mmol) which was used without purification in thesubsequent step.

LCMS (Method E): m/z 350.4 (ES+), at 3.91 min.

¹H NMR: (500 MHz, CD₃OD) δ: ppm 1.43-1.48 (m, 1H), 1.46 (s, 9H),1.87-2.02 (m, 6H), 2.39-2.46 (m, 2H), 2.51-2.58 (m, 1H), 2.76 (br s,2H), 2.81-2.92 (m, 1H), 3.10 (dt, J=12.4, 3.6, 2H), 3.71 (s, 3H), 4.14(dt, J=13.4, 2.4, 2H), 4.59 (br s, 1H), 8.35 (s, 1H)

Step 2) 4N HCl in 1,4-dioxane (16.0 mL, 64.0 mmol) was added to asolution of tert-butyl4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate(2.27 g, 6.50 mmol) in DCM (120 mL) and the mixture stirred at rt for 2h. Concentration in vacuo yielded the title compound (2.23 g) as thehydrochloride salt which was used without further purification.

Data in Table 1.

Alternative synthesis of Intermediate 8,4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine

Step 1) Benzyl 4-oxopiperidine-1-carboxylate (Intermediate 22, 23.6 g,101.1 mmol) and sodium triacetoxyborohydride (28.6 g, 134.8 mmol) wereadded to a solution of 4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine(Intermediate 6, 16.0 g, 96.3 mmol) in DCE (580 mL). After stirring atrt overnight acetic acid (5 mL) was added, and after stirring at rt fora further 2 h benzyl 4-oxopiperidine-1-carboxylate (Intermediate 22,2.35 g, 10.1 mmol) and sodium triacetoxyborohydride (2.86 g, 13.5 mmol)were added. After stirring at rt for 2 h benzyl4-oxopiperidine-1-carboxylate (Intermediate 22, 7.05 g, 30.3 mmol) andsodium triacetoxyborohydride (8.58 g, 40.4 mmol) were added, the mixturewas stirred at rt for 2 h before the addition of benzyl4-oxopiperidine-1-carboxylate (Intermediate 22, 7.05 g, 30.3 mmol) andsodium triacetoxyborohydride (8.58 g, 40.4 mmol). After stirring at rtovernight H₂O (1 L) was added, the phases were separated, and theaqueous phase was extracted with DCM (3×300 mL). The pH of the aqueousphase was adjusted to 7.5 with 6N aqueous NaOH and extracted with DCM(5×300 mL). The combined organic phases were dried (Na₂SO₄), filteredand concentrated in vacuo to yield benzyl4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate(23.2 g, 60.6 mmol).

LCMS (Method A): m/z 384.3 (ES+), at 3.29 min.

¹H NMR: (400 MHz, CD₃OD) δ: ppm 1.40-1.52 (m, 2H), 1.86-2.05 (m, 6H),2.39-2.51 (m, 2H), 2.53-2.66 (m, 1H), 2.75-2.98 (m, 3H), 3.04-3.18 (m,2H), 3.71 (s, 3H), 4.22 (d, J=13.4, 2H), 5.11 (s, 2H), 7.29-7.39 (m,5H), 8.35 (s, 1H)

Step 2) A mixture of 10% palladium on carbon (4.76 g) and benzyl4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate(17.2 g, 44.8 mmol) in EtOH (200 mL) was stirred under an atmosphere ofH₂ (1.5 bar) at rt for 90 min. The mixture was filtered through celite,concentrated in vacuo, re-dissolved in DCM (200 mL) and concentrated invacuo. The re-dissolution and concentration process was repeated fivetimes, and after further drying in vacuo the title compound (10.0 g,40.1 mmol) was obtained as a white solid.

¹H NMR: (400 MHz, DMSO-d₆) δ: ppm 1.40-1.51 (m, 2H), 1.63-1.77 (m, 4H),1.80-1.87 (m, 3H), 2.21-2.32 (m, 2H), 2.35-2.45 (m, 1H), 2.52-2.63 (m,2H), 2.66-2.80 (m, 1H), 2.87-2.99 (m, 2H), 3.09 (d, J=12.5, 2H), 3.59(br s, 3H), 8.31 (s, 1H).

Intermediate 10, 4-(1H-imidazol-2-yl)-1,4′-bipiperidine hydrochloride

Step 1) A solution of 4-(1H-imidazol-2-yl)piperidine hydrochloride(Intermediate 9, 2.30 g, 12.3 mmol), Et₃N (4.96 mL, 35.6 mmol),tert-butyl 4-oxopiperidine-1-carboxylate (Intermediate 7, 2.44 g, 12.2mmol) and ZnCl₂ (84.6 mg, 0.61 mmol) in MeOH (100 mL) was stirred at 60°C. for 5 h. After cooling to rt, sodium cyanoborohydride (3.09 g, 49.2mmol) was added portionwise and the mixture stirred at rt for 16 h.After partitioning between MeOH/DCM (1:9, 300 mL) and H₂O (250 mL) theaqueous phase was extracted with MeOH/DCM (1:9, 300 mL) and the combinedorganic phases were dried (Na₂SO₄) and concentrated in vacuo to yieldtert-butyl 4-(1H-imidazol-2-yl)-1,4′-bipiperidine-1′-carboxylate (5.0 g,colourless oil) which was used without purification in the subsequentstep.

TLC: Rf 0.5 (MeOH/DCM 1:9).

LCMS (Method D): m/z 335.2 (ES+), at 4.95 min.

Step 2) Et₃N (4.16 mL, 29.8 mmol) was added to a solution of tert-butyl4-(1H-imidazol-2-yl)-1,4′-bipiperidine-1′-carboxylate (5.0 g) in DCM(100 mL). After stirring for 15 min di-tert-butyl dicarbonate (4.85 g,22.2 mmol) was added at 0° C. portion wise and the reaction mixturestirred at rt for 16 h. After partitioning between MeOH/DCM (1:9, 300mL) and H₂O (250 mL) the aqueous phase was extracted with MeOH/DCM (1:9,300 mL) and the combined organic phases were dried (Na₂SO₄) andconcentrated in vacuo. Purification by gradient flash chromatography,eluting with 0-3% MeOH in DCM yielded tert-butyl4-[1-(tert-butoxycarbonyl)-1H-imidazol-2-yl]-1,4′-bipiperidine-1′-carboxylate(1.40 g, 3.22 mmol) as a colourless oil.

¹H NMR: (400 MHz, DMSO-d₆) δ: ppm 1.22-1.32 (m, 3H), 1.39 (s, 9H), 1.57(s, 9H), 1.69-1.72 (m, 4H), 1.85-1.91 (m, 2H), 2.20 (br s, 2H), 2.67 (brd, 3H), 2.99 (br s, 2H), 3.96 (dd, J=10.0, 2H), 6.85 (d, J=1.6, 1H),7.42 (d, J=1.2, 1H).

Step 3) 4M HCl in 1,4-dioxane (15 mL, 60 mmol) was added dropwise to asolution of tert-butyl4-[1-(tert-butoxycarbonyl)-1H-imidazol-2-yl]-1,4′-bipiperidine-1′-carboxylate(1.40 g, 3.22 mmol) in 1,4-dioxane (20 mL) at 0° C., and the mixturesubsequently stirred at rt for 4 h. After concentration in vacuotrituration with Et₂O yielded the title compound (1.0 g) as an off-whitesolid.

Data in Table 1.

Intermediate 12, 4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidinehydrochloride

Step 1) A mixture of 4-(4-methyl-1H-imidazol-5-yl)piperidinehydrochloride (Intermediate 11, 2.0 g, 9.9 mmol) and triethylamine (1.7mL, 11.9 mmol) in DMF (40 mL) was stirred at rt. After 5 min tert-butyl4-oxopiperidine-1-carboxylate (Intermediate 7, 2.4 g, 11.9 mmol) andacetic acid (0.68 ml, 11.9 mmol) were added. After 30 min stirring at rtsodium triacetoxyborohydride (2.52 g, 11.9 mmol) was added. Afterstirring at rt overnight the reaction mixture was concentrated in vacuoand the residue partitioned between DCM (100 mL) and saturated aqueousNaHCO₃ (100 ml). The aqueous phase was extracted with DCM (100 mL) andthe combined organic phases were washed with brine and further dried bypassing through a hydrophobic frit. The solvent was removed in vacuo andthe product oil crystallised on standing for 3 d. After trituration withdiethylether/isohexane the solid was collected by filtration and driedin vacuo to yield tert-butyl4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidine-1′-carboxylate (1.64 g,4.7 mmol).

LCMS (Method B): m/z 349.2 (ES+), at 1.24 min.

¹H NMR: (400 MHz, CDCl3) δ: ppm 1.39-1.51 (m, 11H), 1.68-1.91 (m, 6H),2.20 (s, 3H), 2.24-2.37 (m, 2H), 2.40-2.53 (m, 1H), 2.55-2.77 (m, 3H),2.97-3.07 (m, 2H), 4.15 (br s, 2H), 7.45 (s, 1H) (1 exchangeable protonnot observed)

Step 2) 4N HCl in 1,4-dioxane (10.0 mL, 40.0 mmol) was added to asolution of tert-butyl4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidine-1′-carboxylate (1.6 g,4.6 mmol) in DCM (20 mL) and the mixture stirred at rt. After 2 h thereaction mixture was concentrated in vacuo, and the residue taken up andre-evaporated from DCM (×2) to yield the title compound (3.22 g,contains residual solvent) which was used without further purificationin the synthesis of Example 2.

Data in Table 1.

A sample of Intermediate 12 (100 mg) was dissolved in DCM (5 mL) and aminimum amount of MeOH, solid NaCO₃ (200 mg) was added and the mixturestirred for 2 h. The reaction mixture was filtered and the filtrateconcentrated under a flow of N₂ then in vacuo to yield the titlecompound as the free base (31 mg, 0.12 mmol).

LCMS (Method A): m/z 249.3 (ES+), at 2.24 min.

¹H NMR: (400 MHz, CD₃OD) δ: ppm 1.47 (qd, J=12.3, 4.1, 2H), 1.72-1.83(m, 4H), 1.86-1.93 (m, 2H), 2.16 (s, 3H), 2.30-2.39 (m, 2H), 2.46 (tt,J=11.6, 3.5, 1H), 2.53-2.67 (m, 3H), 3.02-3.14 (m, 4H), 7.40 (s, 1H) (2exchangeable protons not observed).

Intermediate 14, 4-(1-propyl-1H-imidazol-2-yl)piperidine hydrochloride

Step 1) Sodium hydride (60% in mineral oil, 478 mg, 12.0 mmol) was addedto a solution of tert-butyl 4-(1H-imidazol-2-yl)piperidine-1-carboxylate(Intermediate 13, 2.50 g, 9.95 mmol) in DMF (50 mL) at 0° C. Afterstirring at 0° C. for 20 min 1-iodopropane (1.16 mL, 11.9 mmol) wasadded and the reaction was stirred at rt for 2 h before partitioningbetween EtOAc (200 mL) and H₂O (150 mL). The aqueous phase was extractedwith EtOAc (200 mL) and the combined organic phases were dried (Na₂SO₄),filtered, and concentrated in vacuo. Purification by gradient flashchromatography, eluting with 0-10% MeOH in DCM yielded tert-butyl4-(1-propyl-1H-imidazol-2-yl)piperidine-1-carboxylate (2.90 g, 9.89mmol) as a yellow oil.

LCMS (Method C): m/z 294.5 (ES⁺), at 1.77 min.

¹H NMR: (400 MHz, DMSO-d₆) δ: ppm 0.81-0.92 (m, 3H), 1.41 (s, 9H),1.51-1.70 (m, 6H), 2.89-2.97 (m, 3H), 3.81-3.88 (m, 2H), 3.97-4.00 (d,J=13.6, 2H), 6.78 (s, 1H), 7.04 (s, 1H).

Step 2) The title compound (2.60 g) was prepared from tert-butyl4-(1-propyl-1H-imidazol-2-yl)piperidine-1-carboxylate (2.90 g, 9.89mmol) and 4M HCl in 1,4-dioxane (15 mL, 60.0 mmol) using the methods ofIntermediate 10, Step 3.

Data in Table 1.

Intermediate 15, 4-(1-propyl-1H-imidazol-2-yl)-1,4′-bipiperidinehydrochloride

The title compound (0.70 g, 1.81 mmol) was prepared from4-(1-propyl-1H-imidazol-2-yl)piperidine hydrochloride (Intermediate 14,1.00 g) and tert-butyl 4-oxopiperidine-1-carboxylate (Intermediate 7,868 mg, 4.36 mmol) using the methods of Intermediate 10, Steps 1 and 3.

Data in Table 1.

Intermediate 17, ethyl5-(1,4′-bipiperidin-4-yl)-4H-1,2,4-triazole-3-carboxylate hydrochloride

The title compound was prepared over two steps from ethyl5-(piperidin-4-yl)-4H-1,2,4-triazole-3-carboxylate (Intermediate 16, 448mg, 2.00 mmol), tert-butyl 4-oxopiperidine-1-carboxylate (Intermediate7, 478 mg, 2.40 mmol), sodium triacetoxyborohydride (610 mg, 2.88 mmol)and acetic acid (137 μL, 2.39 mmol) using the methods of Intermediate12.

Data in Table 1.

Intermediate 19, 4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidinehydrochloride

Step 1) A mixture of 4-(1H-1,2,4-triazol-5-yl)piperidine hydrochloride(Intermediate 18, 377 mg, 2.00 mmol), tert-butyl4-oxopiperidine-1-carboxylate (Intermediate 7, 478 mg, 2.40 mmol),acetic acid (137 μL, 2.39 mmol) and Et₃N (279 μL, 2.00 mmol) was stirredat rt for 30 min before addition of sodium triacetoxyborohydride (610mg, 2.88 mmol) and stirring at rt for 3 d. Further tert-butyl4-oxopiperidine-1-carboxylate (Intermediate 7, 300 mg, 1.51 mmol) andacetic acid (100 μL, 1.75 mmol) were added and the reaction mixture wasstirred at rt for 30 min before addition of sodium triacetoxyborohydride(400 mg, 1.89 mmol) and stirring at rt overnight. After concentration invacuo purification by gradient flash chromatography, eluting with 0-10%MeOH in DCM, followed by 10% (7N NH₃ in MeOH) in DCM yielded tert-butyl4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate (350 mg, 1.04mmol) as a white solid.

LCMS (Method B): m/z 336.2 (ES⁺), at 0.94 min.

¹H NMR: (400 MHz, CD₃OD) δ: ppm, 1.45 (s, 9H), 1.86-1.95 (m, 4H),2.10-2.13 (m, 2H), 2.56-2.61 (m, 2H), 2.69-2.81 (m, 3H), 2.88-2.94 (m,1H), 3.03-3.19 (m, 4H), 4.15-4.18 (m, 2H), 8.16 (s, 1H) (oneexchangeable proton not observed).

Step 2) The title compound (320 mg) was prepared from tert-butyl4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate (350 mg, 1.04mmol) and 4M HCl in 1,4-dioxane (10 mL, 40.0 mmol) in MeOH (10 mL) usingthe methods of Intermediate 12.

Data in Table 1.

Intermediate 21, 4-(5-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidinehydrochloride

Step 1) A mixture of 4-(3-methyl-1H-1,2,4-triazol-5-yl)piperidinedihydrochloride (Intermediate 20, 478 mg, 2.00 mmol), tert-butyl4-oxopiperidine-1-carboxylate (Intermediate 7, 478 mg, 2.40 mmol),acetic acid (137 μL, 2.39 mmol) and Et₃N (558 μL, 4.00 mmol) was stirredat rt for 30 min before addition of sodium triacetoxyborohydride (610mg, 2.88 mmol) and stirring at rt overnight. After concentration invacuo purification by gradient flash chromatography, eluting with 0-10%(7N NH₃ in MeOH) in DCM yielded tert-butyl4-(5-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate (310mg, 0.89 mmol) as a colourless solid.

LCMS (Method B): m/z 350.2 (ES⁺), at 1.01 min.

¹H NMR: (400 MHz, CD₃OD) δ: ppm 1.45 (s, 9H), 1.48-1.55 (m, 2H),1.91-1.99 (m, 7H), 2.11-2.15 (m, 2H), 2.68-2.90 (m, 6H), 3.24-3.27 (m,2H), 4.16-4.20 (m, 2H) (one exchangeable proton not observed).

Step 2) The title compound (240 mg, 0.93 mmol) was prepared fromtert-butyl4-(5-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine-1′-carboxylate (310mg, 0.89 mmol) and 4M HCl in 1,4-dioxane (5 mL, 20.0 mmol) in MeOH (5mL) using the methods of Intermediate 12.

Data in Table 1.

TABLE 1 Intermediates. Intermediate Name Data 13-(piperidin-4-yl)quinolin-2(1H)- Commercially available, CAS No.205058-78-2 one 2 7-fluoro-3-(piperidin-4-yl)quinolin- Commerciallyavailable, 2(1H)-one CAS No. 885654-35-3 (free base), 885609-87-0(hydrochloride salt) 3 (R)-methyl 2-amino-3-(7-methyl- Commerciallyavailable, 1H-indazol-5-yl)propanoate CAS No. 890044-58-3 (free base),CAS No. 1414976-14-9 (dihydrochloride salt) 4(2R)-3-(7-methyl-1H-indazol-5- LCMS (Method A): m/z 474.3 (ES+), at 1.82min. yl)-2-({[4-(2-oxo-1,2- ¹H NMR (400 MHz, DMSO-d₆) δ:dihydroquinolin-3-yl)piperidin-1- 1.25-1.36 (m, 2H), 1.72-1.78 (m, 2H),2.48 (s, 3H), yl]carbonyl}amino)propanoic acid 2.66-2.78 (m, 2H),2.88-2.94 (m, 1H), 2.97-3.03 (m, 1H), 3.10 (dd, J = 8.4, 3.4, 1H), 4.08(d, J = 12.0, 2H), 4.24-4.30 (m, 1H), 6.57 (d, J = 8.0, 1H), 7.04 (s,1H), 7.15 (dd, J = 12.4, 1.2, 1H), 7.27 (d, J = 8.4, 1H), 7.41-7.45 (m,2H), 7.54 (s, 1H), 7.62 (dd, J = 6.8, 1.2, 1H), 7.97 (s, 1H), 11.69 (s,1H), 12.1-13.1 (br s, 2H) 5 (2R)-2-({[4-(7-fluoro-2-oxo-1,2- LCMS(Method A): m/z 490.5 (ES−), dihydroquinolin-3-yl)piperidin-1- 492.2(ES+), at 2.06 min. ¹H NMR (400 MHz, yl]carbonyl}amino)-3-(7-methyl-DMSO-d₆) δ: ppm 1.14-1.44 (m, 2H), 1H-indazol-5-yl)propanoic acid1.65-1.80 (m, 2H), 2.40 (s, 3H), 2.60-2.79 (m, 2H), 2.79-2.90 (m, 1H),2.92-3.12 (m, 2H), 3.58 (t, J = 6.4, 1H), 3.93 (d, J = 12.9, 1H),3.99-4.13 (m, 2H), 6.33 (br s, 1H), 6.90-7.07 (m, 3H), 7.32 (s, 1H),7.60 (s, 1H), 7.69 (dd, J = 9.4, 6.2, 1H), 7.89 (s, 1H), 11.88 (br s,1H), 12.94 (br s, 1H) 6 4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidineCommercially available, CAS No. 297172-18-0 (free base), 297171-80-3(HCl salt) 7 tert-butyl 4-oxopiperidine-1- Commercially available,carboxylate CAS No. 79099-07-3 8 4-(4-methyl-4H-1,2,4-triazol-3-yl)- ¹HNMR (500 MHz, CD₃OD) δ: 2.04-2.17 (m, 1,4′-bipiperidine hydrochloride2H), 2.27-2.38 (m, 2H), 2.39-2.51 (m, 4H), 3.12-3.19 (m, 2H), 3.35-3.43(m, 2H), 3.54-3.75 (m, 4H), 3.78 (d, J = 12.5, 2H), 3.98 (s, 3H), 9.57(s, 1H) (exchangeable protons not observed) 94-(1H-imidazol-2-yl)piperidine Commercially available, CAS No.647024-44-0 (free base), 239800-93-2 (hydrochloride salt) 104-(1H-imidazol-2-yl)-1,4′- LCMS (Method A): m/z 235.3 (ES+), at 0.32min. bipiperidine hydrochloride ¹H NMR (400 MHz, CD₃OD) δ: ppm 1.94-2.20(m, 2H), 2.24-2.53 (m, 6H), 3.03-3.20 (m, 3H), 3.32-3.40 (m, 1H),3.50-3.73 (m, 4H), 3.74-3.83 (m, 2H), 7.54 (s, 2H) (exchangeable protonsnot observed) 11 4-(4-methyl-1H-imidazol-5-yl)piperidine Commerciallyavailable, CAS No. 155511-82-3 (free base), 1246551-65- 4 (hydrochloridesalt) 12 4-(4-methyl-1H-imidazol-5-yl)- LCMS (Method A): m/z 249.3(ES+), at 2.15 min. 1,4′-bipiperidine hydrochloride ¹H NMR: (400 MHz,CD₃OD) δ:ppm 2.05-2.26 (m, 4H), 2.22-2.35 (m, 2H), 2.38 (s, 3H),2.45-2.56 (m, 2H), 3.05-3.24 (m, 3H), 3.26-3.40 (m, 3H), 3.56-3.79 (m,4H), 8.76 (s, 1H) (exchangeable protons not observed) 13 tert-butyl4-(1H-imidazol-2- Commercially available, yl)piperidine-1-carboxylateCAS No. 158654-96-7 14 4-(1-propyl-1H-imidazol-2- LCMS (Method A): m/z194.2 (ES+), at 1.91 min. yl)piperidine hydrochloride ¹H NMR (400 MHz,DMSO-d₆ + D₂O) δ: ppm 0.86 (t, J = 7.3, 3H), 1.70-1.84 (m, 2H),1.92-2.12 (m, 4H), 3.01-3.25 (m, 2H), 3.38 (d, J = 12.5, 2H), 3.54 (t, J= 12.1, 1H), 4.11 (t, J = 7.5, 2H), 7.58 (d, J = 2.1, 1H), 7.63 (d, J =12.1, 1H) (exchangeable protons not observed) 154-(1-propyl-1H-imidazol-2-yl)-1,4′- LCMS (Method A): m/z 277.3 (ES+), at2.81 min. bipiperidine hydrochloride ¹H NMR (400 MHz, CD₃OD) δ: ppm 1.04(t, J = 7.4, 3H), 1.88-2.00 (m, 2H), 2.08-2.20 (m, 2H), 2.28-2.44 (m,4H), 2.52 (br d, J = 15.8, 2H), 3.06-3.26 (m, 3H), 3.40-3.50 (m, 2H),3.57-3.84 (m, 5H), 4.26 (dd, J = 7.8, 7.0, 2H), 7.59 (d, J = 2.3, 1H),7.63 (d, J = 2.3, 1H) (exchangeable protons not observed) 16 ethyl5-(piperidin-4-yl)-4H-1,2,4- Commercially available,triazole-3-carboxylate CAS No. 1513304-93-2 (free base), 1795504- 00-5(hydrochloride salt) 17 ethyl 5-(1,4′-bipiperidin-4-yl)-4H- LCMS (MethodB): m/z 308.2 (ES+), at 0.20 min. 1,2,4-triazole-3-carboxylate ¹H NMR(400 MHz, CD₃OD) δ: ppm hydrochloride 1.41 (t, J = 7.1, 3H), 1.99-2.25(m, 2H), 2.39-2.46 (m, 4H), 3.07-3.18, (m, 3H), 3.29-3.31 (m, 1H)3.32-3.38 (m, 2H), 3.58-3.68 (m, 4H), 3.71-3.76 (m, 2H), 4.45 (q, J =7.1, 2H) (exchangeable protons not observed) 184-(1H-1,2,4-triazol-5-yl)piperidine Commercially available, CAS No.893424-25-4 (free base), 1417359-91- 1 (hydrochloride salt) 194-(4H-1,2,4-triazol-3-yl)-1,4′- LCMS (Method B): m/z 236.2 (ES+), at0.50 min. bipiperidine hydrochloride ¹H NMR (400 MHz, CD₃OD) δ: ppm2.08-2.17 (m, 2H), 2.22-2.32 (m, 2H), 2.44-2.54 (m, 4H), 3.11-3.18 (m,3H), 3.36-3.47 (m, 2H), 3.60-3.67 (m, 3H), 3.75-3.82 (m, 2H), 9.35 (s,1H) (exchangeable protons not observed) 204-(3-methyl-1H-1,2,4-triazol-5-yl)piperidine Commercially available, CASNo. 933713-90-7 (free base), 1221724-59- 9 (dihydrochloride salt) 214-(5-methyl-4H-1,2,4-triazol-3-yl)- LCMS (Method B): m/z 250.2 (ES+), at1,4′-bipiperidine hydrochloride 0.68 min. ¹H NMR (400 MHz, CD₃OD) δ: ppm2.02-2.14 (m, 2H), 2.21-2.31 (m, 2H), 2.42-2.49 (m, 4H), 2.68 (s, 3H),3.09-3.22 (m, 3H), 3.31-3.40 (m, 2H), 3.58-3.67 (m, 3H), 3.70-3.78 (m,2H) (exchangeable protons not observed) 22 benzyl 4-oxopiperidine-1-Commercially available, CAS No. 19099-93-5 carboxylate

SYNTHESIS OF EXAMPLES Typical Procedures for the Preparation of ExamplesVia Amide Coupling, as Exemplified by the Preparation of the BelowExamples Procedure 1 Example 2,N-[(2R)-1-[4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidin-r-yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide

A mixture of 4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidinehydrochloride (Intermediate 12, contaminated with solvent residues,assumed to be 21.0 mmol),(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoicacid (Intermediate 4, 9.93 g, 21.0 mmol), HATU (8.00 g, 20.9 mmol) andDIPEA (14.6 mL, 83.8 mmol) in DMF (150 mL) was stirred at roomtemperature overnight before concentration in vacuo. Purification bygradient flash chromatography, eluting with 0-100% solvent B in DCM(where solvent B is 7N NH₃ in MeOH/DCM, 1:9) yielded the title compound(7.50 g, 10.7 mmol) as a white solid.

Data in Table 2.

Example 5,4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)-N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}piperidine-1-carboxamide

The title compound (45 mg, 0.06 mmol) was prepared from(2R)-2-({[4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)-3-(7-methyl-1H-indazol-5-yl)propanoicacid (Intermediate 5, 74 mg, 0.15 mmol),4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidine hydrochloride(Intermediate 8, 97 mg), HATU (69 mg, 0.18) and Et₃N (0.21 mL, 1.51mmol) in DMF (1.5 mL) using methods of Example 4. The title compound waspurified by gradient flash column chromatography eluting with 0-100%solvent B in DCM (where solvent B is 7N NH₃ in MeOH/DCM, 1:9), followedby preparative reversed phase HPLC (Phenomenex Gemini-NX 5 μm C18column, 100×30 mm, eluting with 10 to 40% MeCN/Solvent B over 12.5 minat 30 mL/min [where solvent B is 0.2% of (28% NH₃/H₂O) in H₂O] andcollecting fractions by monitoring at 205 nm).

Data in Table 2.

Procedure 2 Example 9,5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylicacid

Lithium hydroxide monohydrate (9 mg, 0.21 mmol) was added to a solutionof ethyl5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylate(Example 6, 110 mg, 0.14 mmol) in MeOH (10 mL) and H₂O (2 mL) and thereaction mixture was stirred at rt overnight. Further lithium hydroxidemonohydrate (10 mg, 0.24 mmol) was added and after stirring at rt for 1d the mixture was partially concentrated in vacuo to remove the MeOH. 1Naqueous HCl was added to the residue, which was then concentrated invacuo and purified by preparative reversed phase HPLC (PhenomenexGemini-NX 5 μm C18 column, 100×30 mm, eluting with 5 to 35% MeCN/SolventB over 12.5 min at 30 mL/min [where solvent B is 0.2% of (28% NH₃/H₂O)in H₂O] and collecting fractions by monitoring at 205 nm) to yield thetitle compound (10 mg, 0.01 mmol) as a colourless solid.

Data in Table 2.

Further examples prepared by the above procedures are detailed in Table2.

TABLE 2 Ex Intermediates/ LCMS data No. Name Procedure ¹H NMR (Method A)1 N-[(2R)-1-[4-(1H- 4, 10 (400 MHz, DMSO-d₆) δ: ppm m/z 688.6 (ES⁻),imidazol-2-yl)-1,4′- Procedure 1 −0.07-0.09 (m, 1H), 0.61 (d, J = 8.6,1H), 690.4 (ES⁺) at bipiperidin-1′-yl]-3-(7- 1.08-1.92 (m, 10H),2.00-2.35 (m, 5H), 2.70 min, 100% methyl-1H-indazol-5-yl)- 2.68-2.81 (m,3H), 2.81-2.95 (m, 3H), 1-oxopropan-2-yl]-4-(2- 2.96-3.07 (m, 1H), 3.30(s, 3H), 3.83-4.02 (m, 1H), oxo-1,2-dihydroquinolin- 4.05-4.23 (m, 2H),4.35 (d, J = 11.7, 1H), 3-yl)piperidine-1- 4.76 (q, J = 7.4, 1H), 4.84(q, J = 7.8, 1H), carboxamide 6.70 (d, J = 7.0, 1H), 6.83 (br s, 2H),7.01 (s, 1H), 7.07-7.19 (m, 1H), 7.20-7.31 (m, 1H), 7.35 (s, 1H),7.37-7.48 (m, 2H), 7.56-7.72 (m, 2H), 7.96 (s, 1H), 11.77 (s, 1H), 13.01(s, 1H) 2 N-[(2R)-1-[4-(4-methyl- 4, 12 (400 MHz, CD₃OD) δ: ppm −0.30 tom/z 704.7 (ES⁺) 1H-imidazol-5-yl)-1,4′- Procedure 1 −0.27 (m, 1H),0.70-0.77 (m, 1H), at 3.69 min, bipiperidin-1′-yl]-3-(7- 1.24-1.32 (m,1H), 1.42-1.94 (m, 11H), 100% methyl-1H-indazol-5-yl)- 2.17 (s, 3H),2.30-2.57 (m, 7H), 2.84-3.12 (m, 1-oxopropan-2-yl]-4-(2- 7H), 3.94-4.10(m, 1H), 4.18-4.25 (m, oxo-1,2-dihydroquinolin- 2H), 4.52 (d, J = 13.0,1H), 5.02-5.08 (m, 3-yl)piperidine-1- 1H), 7.16 (s, 1H), 7.22-7.26 (m,1H), carboxamide 7.30-7.32 (m, 1H), 7.41 (s, 1H), 7.44-7.51 (m, 2H),7.63-7.67 (m, 1H), 7.75 (s, 1H), 8.01 (s, 1H) (4 exchangeable protonsnot observed) 3 N-{(2R)-3-(7-methyl-1H- 4, 15 (400 MHz, CD₃OD) δ: ppm−0.35 to m/z 732.6 (ES⁺) indazol-5-yl)-1-oxo-1-[4- Procedure 1 −0.30 (m,1H), 0.77-0.83 (m, 1H), 0.96 (t, at 3.41 min, (1-propyl-1H-imidazol- J =7.4, 3H), 1.23-2.12 (m, 14H), 95% 2-yl)-1,4′-bipiperidin-1′- 2.31-2.42(m, 4H), 2.54-2.58 (m, 3H), yl]propan-2-yl}-4-(2- 2.86-3.13 (m, 7H),3.91-3.94 (m, 3H), oxo-1,2-dihydroquinolin- 4.21-4.25 (m, 2H), 4.49-4.52(m, 1H), 3-yl)piperidine-1- 5.04-5.05 (m, 1H), 6.85 (s, 1H), 6.99 (s,1H), carboxamide 7.15 (s, 1H), 7.22-7.25 (m, 1H), 7.31-7.33 (m, 1H),7.44-7.51 (m, 2H), 7.65-7.66 (m, 1H), 7.75 (s, 1H), 8.00 (s, 1H) (3exchangeable protons not observed) 4 N-{(2R)-3-(7-methyl-1H- 4, 8  (400MHz, CD₃OD) δ: ppm −0.30 to m/z 705.8 (ES⁺) indazol-5-yl)-1-[4-(4-Procedure 1 −0.25 (m, 1H), 0.82-0.86 (m, 1H), at 3.34 min,methyl-4H-1,2,4-triazol- 1.23-2.04 (m, 13H), 2.33-2.45 (m, 3H), 100%3-yl)-1,4′-bipiperidin-1′- 2.57 (s, 3H), 2.65-3.11 (m, 7H), 3.73 (s,3H), yl]-1-oxopropan-2-yl}-4- 3.94-3.97 (m, 1H), 4.16-4.24 (m, 2H),(2-oxo-1,2- 4.50-4.53 (m, 1H), 5.02-5.08 (m, 1H), dihydroquinolin-3-7.16 (s, 1H), 7.21-7.25 (m, 1H), 7.32 (d, yl)piperidine-1- J = 7.8, 1H),7.45-7.50 (m, 2H), carboxamide 7.63-7.65 (m, 1H), 7.73 (s, 1H), 8.00 (s,1H), 8.36 (s, 1H) (3 exchangeable protons not observed) 54-(7-fluoro-2-oxo-1,2- 5, 8  (400 MHz, CD₃OD) δ: ppm −0.45 to m/z 723.6(ES⁺), dihydroquinolin-3-yl)-N- Procedure 1 −0.23 (m, 1H), 0.61-0.79 (m,1H), at 3.07 min, {(2R)-3-(7-methyl-1H- 1.12-1.64 (m, 4H), 1.64-2.00 (m,8H), 2.07 (d, 100% indazol-5-yl)-1-[4-(4- J = 11.3, 1H), 2.27-2.62 (m,6H), 2.67 (d, methyl-4H-1,2,4-triazol- J = 9.4, 1H), 2.76-3.15 (m, 6H),3-yl)-1,4′-bipiperidin-1′- 3.61-3.79 (m, 3H), 3.96 (d, J = 13.7, 1H),4.22 (d, yl]-1-oxopropan-2- J = 13.3, 2H), 4.51 (d, J = 12.9, 1H),yl}piperidine-1- 5.00-5.14 (m, 1H), 6.94-7.09 (m, 2H), 7.16 (s,carboxamide 1H), 7.38-7.50 (m, 1H), 7.57-7.78 (m, 2H), 7.94-8.06 (m,1H), 8.30-8.42 (m, 1H) (3 exchangeable protons not observed). 6 ethyl5-{1′-[(2R)-3-(7- 4, 17 (400 MHz, CD₃OD) δ: ppm −0.33 to m/z 763.7 (ES⁺)methyl-1H-indazol-5-yl)- Procedure 1 −0.29 (m, 1H), 0.83-0.87 (m, 1H),at 3.26 min, 2-({[4-(2-oxo-1,2- 1.20-2.46 (m, 16H), 2.57 (s, 3H), 95%dihydroquinolin-3- 2.66-3.11 (m, 7H), 3.92-3.99 (m, 1H), yl)piperidin-1-4.21-4.24 (m, 2H), 4.27-40 (m, 2H), 4.42-4.70 (m,yl]carbonyl}amino)propanoyl]- 4H), 5.00-5.08 (m, 1H), 7.17 (s, 1H),1,4′-bipiperidin-4- 7.22-7.25 (m, 1H), 7.31-7.33 (m, 1H),yl}-4H-1,2,4-triazole-3- 7.42-7.51 (m, 2H), 7.65-7.66 (m, 1H),carboxylate 7.75 (s, 1H), 8.02 (s, 1H) (4 exchangeable protons notobserved) 7 N-{(2R)-3-(7-methyl-1H- 4, 19 (400 MHz, CD₃OD) δ: ppm −0.29to m/z 691.6 (ES⁺) indazol-5-yl)-1-oxo-1-[4- Procedure 1 −0.19 (m, 1H),0.83-0.86 (m, 1H), at 2.92 min, (4H-1,2,4-triazol-3-yl)- 1.48-1.59 (m,3H), 1.82-2.07 (m, 5H), 100% 1,4′-bipiperidin-1′- 2.24-2.35 (m, 2H),2.46-2.60 (m, 5H), yl]propan-2-yl}-4-(2- 2.63-2.71 (m, 1H), 2.87-3.17(m, 7H), oxo-1,2-dihydroquinolin- 3.23-3.41 (m, 2H), 3.59-3.63 (m, 1H),3-yl)piperidine-1- 4.08-4.24 (m, 2H), 4.63-4.67 (m, 1H), carboxamide7.10-7.12 (m, 1H), 7.22-7.25 (m, 1H), 7.32 (d, J = 8.2, 1H), 7.47-7.51(m, 2H), 7.60-7.64 (m, 1H), 7.73 (s, 1H), 8.07 (s, 1H), 8.58-8.61 (m,1H) (4 exchangeable protons not observed, 2 protons obscured by solventpeak) 8 N-{(2R)-3-(7-methyl-1H- 4, 21 (400 MHz, CD₃OD) δ: ppm −0.23 tom/z 705.6 (ES⁺) indazol-5-yl)-1-[4-(5- Procedure 1 −0.18 (m, 1H),0.83-0.87 (m, 1H), at 3.00 min, methyl-4H-1,2,4-triazol- 1.52-1.75 (m,2H), 1.80-2.10 (m, 5H) 95% 3-yl)-1,4′-bipiperidin-1′- 2.22-2.38 (m, 2H),2.41-2.60 (m, 8H), yl]-1-oxopropan-2-yl}-4- 2.65-2.71 (m, 1H), 2.80-3.10(m, 8H) (2-oxo-1,2- 3.15-3.25 (m, 1H), 3.55-3.70 (m, 2H),dihydroquinolin-3- 4.06-4.24 (m, 3H), 4.62-4.65 (m, 1H),yl)piperidine-1- 5.00-5.03 (m, 1H), 7.21-7.23 (m, 2H), carboxamide7.31-7.34 (m, 1H), 7.46-7.51 (m, 2H), 7.64-7.72 (m, 2H), 7.96-8.05 (m,1H) (4 exchangeable protons not observed) 9 5-{1′-[(2R)-3-(7-methyl-Example 6 (400 MHz, CD₃OD) δ: ppm −0.33 to m/z 735.7 (ES⁺)1H-indazol-5-yl)-2-({[4- Procedure 2 −0.27 (m, 1H), 0.69-0.75 (m, 1H),at 2.33 min, (2-oxo-1,2- 1.22-1.44 (m, 2H), 1.51-1.82 (m, 4H), 100%dihydroquinolin-3- 1.90-2.46 (m, 7H), 2.50-2.60 (m, 4H), yl)piperidin-1-2.62-3.17 (m, 7H), 3.50-3.70 (m, 2H), yl]carbonyl}amino)propanoyl]-3.94-4.24 (m, 3H), 4.52-4.65 (m, 1H), 1,4′-bipiperidin-4- 5.06-5.09 (m,1H), 7.15 (s, 1H), 7.21-7.26 (m, yl}-4H-1,2,4-triazole-3- 1H), 7.31-7.33(m, 1H), 7.47-7.51 (m, carboxylic acid 2H), 7.65-7.69 (m, 1H), 7.75 (s,1H), 7.97-8.02 (m, 1H) (5 exchangeable protons not observed)Biological and Biophysical Methods

Cloning, Baculovirus Generation, Large-Scale Infection of Sf21 Cells andMembrane Preparation.

Human Calcitonin Receptor Like Receptor (CRLR) and human RAMP1 werecloned into Invitrogen's (ThermoFisher Scientific, UK) pFastBac dualexpression vector. Transposition of CRLR/RAMP1 DNA was performed usingInvitrogen's Bac-to-Bac Baculovirus Expression Systems. P0 baculoviruswas generated by transfecting SF9 cells with bacmid DNA usingCellfectin® II transfection reagent (ThermoFisher Scientific, UK,catalog number 10362-100). Following P0 generation P1 virus was thengenerated ready for large scale infection and membrane preparation. Sf21cells were grown in expression medium ESF921 (Expression Systems, USA,catalog number 96-001-01) supplemented with 10% heat-inactivated FBS and1% Pen/Strep and were infected at a cell density of 2.5×10⁶ cells/mL andan MOI of 2. Expression was carried out over 48 h in a shaking incubatorset at 27° C. The cell culture was centrifuged at 2,500 rcf for 10 minat 4° C. The pellets were resuspended in cold PBS supplemented withRoche's Complete EDTA-free protease inhibitor cocktail tablets (RocheApplied Sciences, catalog number 05056489001), 1 mM PMSF and 1 mM EDTA.The resuspended cell paste was then centrifuged at 3,273 rcf for 12 minat 4° C. The supernatant was discarded and the pellet frozen at −80° C.The cell pellet from a 4 L culture was resuspended in buffer containing50 mM Hepes pH 7.5, 150 mM NaCl, 8 Roche EDTA-free protease inhibitorcocktail tablets and 1 mM PMSF. The suspension was left stirring at rtfor 1 h and then homogenised for 90 s at 9,500 rpm using a VDI 25 (VWR,USA) homogeniser. The cells were then lysed using a Microfluidizerprocessor M-110L Pneumatic (Microfluidics, USA). After lysis, themixture was homogenised for 90 s at 9,500 rpm and then centrifuged at335 rcf for 10 min. The supernatant was then further ultra-centrifugedat 42,000 rpm for 90 min. After ultra-centrifugation, the supernatantwas discarded and the pellet was resuspended in 50 mL (25 mL for each 2L culture) of buffer containing 50 mM Hepes pH 7.5, 150 mM NaCl, 3 RocheEDTA-free protease inhibitor cocktail tablets and 1 mM PMSF. Thesuspension was then homogenised for 90 s at 9,500 rpm. The resultingmembranes were then stored at −80° C.

Radioligand Binding Assay.

Human CGRP receptors (consisting of CRLR and RAMP1) expressed in insectSf21 cell membrane homogenates were re-suspended in the binding buffer(10 mM HEPES, pH 7.4, 5 mM MgCl₂, 0.2% BSA) to a final assayconcentration of 0.6 μg protein per well. Saturation isotherms weredetermined by the addition of various concentrations of ³H-telcagepant(Ho et al, The Lancet, 2008, 372, 2115) (in a total reaction volume of250 μL) for 60 min at room temperature. At the end of the incubation,membranes were filtered onto a unifilter, a 96-well white microplatewith bonded GF/B filter pre-incubated with 0.5% PEI, with a Tomtec cellharvester and washed 5 times with distilled water. Non-specific binding(NSB) was measured in the presence of 10 nM MK-3207 hydrochloride (CASNo. 957116-20-0). Radioactivity on the filter was counted (1 min) on amicrobeta counter after addition of 50 μL of scintillation fluid. Forinhibition experiments, membranes were incubated with 0.5 nM³H-telcagepant and 10 concentrations of the inhibitory compound(0.001-10 μM). IC₅₀ values were derived from the inhibition curve andthe affinity constant (K₁) values were calculated using theCheng-Prussoff equation (Cheng et al, Biochem. Pharmacol. 1973, 22,3099-3108). The pK_(i) values (where pK_(i)=−log₁₀ K_(i)) of certaincompounds of the invention are tabulated below.

cAMP Functional Assay.

cAMP production following receptor activation was determined using theHomogeneous Time-Resolved Fluorescence (HTRF) cAMP dynamic-2 assay(Cisbio, France). The human neuroblastoma cell line SK-N-MC endogenouslyexpressing the human CGRP receptor was seeded at a density of 12,500cells/well in solid walled 96 well half area plates (Costar, CatalogNumber 3688, Corning Life Sciences, Germany). After 16 h incubation at37° C. media was removed and cells were incubated at 37° C. for 30 minin serum free media containing 500 μM IBMX (Tocris, Abingdon, UK,Catalog Number 2845) and increasing concentrations of test antagonist.Following this cells were challenged with an EC₈₀ concentration of humanCGRP (0.3 nM) for a further 30 min at 37° C. and then cAMP productionwas determined as manufacturer's instructions before plates were read ona PheraStar fluorescence plate reader (BMG LabTech, Germany). IC₅₀values were derived from the inhibition curve. The pIC₅₀ values (wherepIC₅₀=−log₁₀ IC₅₀) were converted to a functional pK_(b) value using amodified Cheng-Prussoff equation where K_(d)=agonist EC₅₀ and Lhot=agonist challenge concentration. The pK_(b) values of certaincompounds of the invention are detailed in Table 3.

TABLE 3 Ex pK_(i) pK_(b) No. Name Structure average average 1N-[(2R)-1-[4-(1H-imidazol-2-yl)- 1,4′-bipiperidin-1′-yl]-3-(7-methyl-1H-indazol-5-yl)-1- oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1- carboxamide

10.3 9.7 2 N-[(2R)-1-[4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidin-1′- yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2- dihydroquinolin-3-yl)piperidine-1-carboxamide

10.1 9.5 3 N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-[4-(1-propyl-1H- imidazol-2-yl)-1,4′-bipiperidin-1′-yl]propan-2-yl}-4-(2-oxo-1,2- dihydroquinolin-3-yl)piperidine-1-carboxamide

 9.8 9.4 4 N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4- triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo- 1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide

10.0 9.2 5 4-(7-fluoro-2-oxo-1,2- dihydroquinolin-3-yl)-N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4- (4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1- oxopropan-2-yl}piperidine-1- carboxamide

 9.9 9.2 6 ethyl 5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2- dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]- 1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylate

10.2 9.7 7 N-{(2R)-3-(7-methyl-1H-indazol- 5-yl)-1-oxo-1-[4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′- yl]propan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1- carboxamide

 9.8 9.5 8 N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(5-methyl-4H-1,2,4- triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo- 1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide

10.0 9.5 9 5-{1′-[(2R)-3-(7-methyl-1H- indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1- yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4- triazole-3-carboxylic acid

10.3 9.4

Pharmacokinetic Profiling.

The pharmacokinetic profiles of Examples and reference compounds havebeen assessed in male Sprague Dawley® rats via intravenous (iv),sub-cutaneous (sc) and intranasal (IN) routes of delivery, and in maleCynomolgus Monkeys via iv and sc routes of delivery. Pharmacokineticdata for Examples of the invention and a reference compound, olcegepant,are detailed in Tables 4 and 5.

Methods:

For rat studies, groups of three male Sprague Dawley® rats, typicallyranging in weight between 180 and 300 g, were given a single dose ofExample or reference compound via one of the following routes: iv, sc orIN, using doses, dose volumes and vehicles specified in Table 5. Priorto IN dosing rats were anaesthetised with an intramuscular dose of 25-30mg/kg ketamine cocktail (ketamine, xylazine hydrochloride andacepromazine maleate in saline) and the dose is introduced over 20-30 svia a polyethylene PE-10 tube inserted approximately 5 mm into the nasalcavity of the rat.

For cynomolgus monkey studies, groups of three male monkeys, typicallyranging in weight between 3.0 and 4.5 kg, were given a single dose ofExample or reference compound via one of the following routes: iv or sc,using doses, dose volumes and vehicles specified in Table 5. Followingdosing by the routes above blood samples were taken at several timepoints (typically pre-dose, 0.083, 0.25, 0.5 1, 2, 4, 8 and 24 h) viaserial tail vein bleeds (rat) or cephalic or saphenous vein (monkey)from the animal and centrifuged to separate plasma for analysis byLC/MS/MS assay. WinNonlin v6.2 statistics software (PharsightCorporation, California, USA) was used to generate pharmacokineticparameters using the non-compartmental model.

TABLE 4 Rat iv pharmacokinetics Dose Dose volume Clearance (mg/kg)(mL/kg) Vehicle (mL/min/kg) olcegepant 5 1 10% DMAC + 10% 18SolutolHS15 + 80% Saline Example 2 2 1 10% DMAC + 10% 18 SolutolHS15 +80% Saline Example 4 2 1 10% DMAC + 10% 2 SolutolHS15 + 80% Saline Ratsc pharmacokinetics Dose Dose volume Bioavailability (mg/kg) (mL/kg)Vehicle (%) olcegepant 1 5 10% DMAC + 10% 48 SolutolHS15 + 80% SalineExample 2 1 2 Acidified saline 69 Example 4 2 5 10% DMAC + 10% 82SolutolHS15 + 80% Saline Rat IN pharmacokinetics Dose Doseconcentration, Bioavailability (mg/kg) Dose volume Vehicle (%)olcegepant 1.3  6 mg/mL, 50 μL Acidified saline 8 Example 2 1 12 mg/mL,25 μL Acidified saline 25 Example 4 1 12 mg/mL, 25 μL Acidified saline27

TABLE 5 Cynomolgus monkey iv pharmacokinetics Dose Dose volume Clearance(mg/kg) (mL/kg) Vehicle (mL/min/kg) Example 2 0.5 0.5 Acidified saline24 Example 4 0.5 0.5 Acidified saline 15 Cynomolgus monkey scpharmacokinetics Dose Dose volume Bioavailability (mg/kg) (mL/kg)Vehicle (%) Example 2 0.5 1 Acidified saline 100 Example 4 0.5 1Acidified saline 100

Thermodynamic Solubility Profiling.

A 50 mM DMSO stock solution of test compound was prepared, and fromthis, a working solution of 1 mM was prepared by dilution with DMSO. TheUV absorbance of working solution was scanned from 220 nm to 1000 nm toidentify the wavelength maxima of test compound. The 1 mM workingsolution was then serially diluted in DMSO to different concentrationsto determine linearity/calibration curve. To ascertain the aqueousthermodynamic solubility of test compound, samples were added to avolume of PBS buffer (pH 7.4) or Sodium Phosphate Buffer (pH 6.0) whichwas appropriate to generate a final concentration of 1 mg/mL if all testcompound dissolved. The resulting solution was then kept on a RotoSpinshaker at 50 rpm for 24 h at rt before the solution was filtered using0.45 micron PVDF injector filters in order to remove the insolublefraction of the compound. Subsequently, 150 uL of the filtrate is takenfor quantification using a UV spectrophotometer, acquiring the opticaldensity of standard solutions and test compound at the same wavelengthmaxima. From the optical density of test compound the thermodynamicsolubility is calculated using the linearity/calibration curve andexpressed as micromolar (μM). Solubility profiles of certain compoundsof the invention are detailed in Table 6.

TABLE 6 Thermodynamic Reference Cpd/ solubility (μM) Example pH 6 pH 7.4olcegepant 150 431 Example 1 Not tested Not tested Example 2 2041 18Example 3 1249 2 Example 4 1587 32 Example 5 1541 239 Example 6 Nottested Not tested Example 7 785 342 Example 8 1217 359 Example 9 Nottested Not tested

The invention claimed is:
 1. A method for treating a CGRP-mediatedcerebrovascular or vascular disorder in a subject comprisingadministering to the subject a compound of formula (I):

or a salt thereof or an optical isomer thereof, wherein R¹ is H or F andAr¹ is an optionally substituted 5 membered heterocyclic ring containingat least two nitrogen atoms.
 2. The method of claim 1, wherein thecerebrovascular or vascular disorder is migraine without aura, chronicmigraine, pure menstrual migraine, menstrually-related migraine,migraine with aura, familial hemiplegic migraine, sporadic hemiplegicmigraine, basilar-type migraine, cyclical vomiting, abdominal migraine,benign paroxysmal vertigo of childhood, retinal migraine, statusmigrainosus, cluster headache, dialysis headache, paroxysmal hemicrania,osteoarthritis, hot flashes associated with menopause or medicallyinduced menopause due to surgery or drug treatment, hemicrania continua,cyclic vomiting syndrome, allergic rhinitis, rosacea, dental pain,earache, middle ear inflammation, sunburn, joint pain associated withosteoarthritis and rheumatoid arthritis, cancer pain, fibromyalgia,diabetic neuropathy, pain associated with inflammatory boweldisease—Crohn's disease, gout, complex regional pain syndrome, Behçet'sdisease, endometriosis pain, back pain, or cough.
 3. The method of claim1 wherein the compound is administered via a non-oral route.
 4. Themethod of claim 3 wherein the non-oral route of administration is anintranasal route, a sub-cutaneous route or an intravenous route.
 5. Themethod of claim 1 wherein Ar¹ is an optionally substituted five-memberedheterocyclic ring including at least two nitrogen atoms, wherein theoptional substituents are selected from (C₁-C₆)alkyl and CO₂R² whereinR² is H or (C₁-C₃)alkyl.
 6. The method of claim 1 wherein R¹ is H. 7.The method of claim 1 wherein Ar¹ is a five-membered heterocyclic ringincluding two or three nitrogen atoms, optionally substituted with(C₁-C₆)alkyl.
 8. The method of claim 1 wherein Ar¹ is


9. The method of claim 1, wherein Ar¹ is


10. The method of claim 1, wherein Ar¹ is


11. The method of claim 1, wherein the compound is selected from thegroup consisting of:N-[(2R)-1-[4-(1H-imidazol-2-yl)-1,4′-bipiperidin-1′-yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-[(2R)-1-[4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidin-1′-yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-[4-(1-propyl-1H-imidazol-2-yl)-1,4′-bipiperidin-1′-yl]propan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)-N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}piperidine-1-carboxamide;ethyl5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylate;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-[4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]propan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(5-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;and5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylicacid; or a salt thereof or an optical isomer thereof.
 12. The method ofclaim 1, wherein the compound is selected from the group consisting of:N-[(2R)-1-[4-(1H-imidazol-2-yl)-1,4′-bipiperidin-1′-yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-[(2R)-1-[4-(4-methyl-1H-imidazol-5-yl)-1,4′-bipiperidin-1′-yl]-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl]-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-[4-(1-propyl-1H-imidazol-2-yl)-1,4′-bipiperidin-1′-yl]propan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;4-(7-fluoro-2-oxo-1,2-dihydroquinolin-3-yl)-N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(4-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}piperidine-1-carboxamide;ethyl5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylate;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-[4-(4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]propan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;N-{(2R)-3-(7-methyl-1H-indazol-5-yl)-1-[4-(5-methyl-4H-1,2,4-triazol-3-yl)-1,4′-bipiperidin-1′-yl]-1-oxopropan-2-yl}-4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidine-1-carboxamide;and5-{1′-[(2R)-3-(7-methyl-1H-indazol-5-yl)-2-({[4-(2-oxo-1,2-dihydroquinolin-3-yl)piperidin-1-yl]carbonyl}amino)propanoyl]-1,4′-bipiperidin-4-yl}-4H-1,2,4-triazole-3-carboxylicacid.
 13. The method of claim 1, wherein the compound is:

or a salt thereof or an optical isomer thereof.
 14. The method of claim1, wherein the compound is:


15. The method of claim 1 wherein the compound is

or a salt thereof or an optical isomer thereof.
 16. The method of claim1 wherein the compound is